马钱子碱对体外软骨细胞凋亡线粒体途径的影响  

The Effect of Trychnine on Apoptosis in Chondtocyte of In-vitro Through Mitochondrial Pathway

作　　者：高萌萌[1] 张梅[2] 

机构地区：[1]安徽医科大学 [2]安徽医科大学附属省立医院中医科

出　　处：《中国中医骨伤科杂志》2007年第12期1-4,共4页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：安徽省教育厅自然科学研究项目(2004BA721A01Z15)

摘　　要：目的:探讨中药马钱子主要成分马钱子碱(strychnine)影响体外SD大鼠软骨细胞凋亡的作用机制,为骨关节炎的临床用药提供理论依据。方法:将体外培养的SD大鼠软骨细胞分为5组,分别加入正常培养液、凋亡诱导剂(SNAP)、SNAP+高剂量马钱子碱、SNAP+中剂量马钱子碱、SNAP+低剂量马钱子碱混合培养。并于培养20h后,流式细胞仪检测软骨细胞早期、晚期凋亡率;免疫组化及Western免疫印迹法观察与细胞凋亡相关蛋白表达的变化。结果:马钱子碱可抑制软骨细胞凋亡,其凋亡率随马钱子碱剂量增大而降低;各剂量组早期凋亡率明显高于晚期凋亡率,与晚期凋亡率相比(P<0.05),差异有统计学意义。正常对照组胞浆内细胞色素C(cytochrome C,CytC)及Smac极少表达,SNAP诱导凋亡后,释放入胞浆的CytC及Smac表达明显增加,但随马钱子碱剂量的增大,其活性降低。结论:马钱子碱可抑制SNAP诱导的鼠软骨细胞凋亡,其保护软骨的作用可能是通过线粒体通路实现。Objective： To observe the effective mechanism of stychnine on chondrocyte apoptosis of SD rat induced in vitro, and to provide the theory basis for clinical application of osteoarthritis. Methods： The chondrocytes from SD rats were divided into 5 groups,which were respectively given normal cultured fluid, SNAP-Which is apoptotic inducer, SNAP＋highdose, medium-dose and low-dose strychnine. After 20 hours , Annexin-V/PI was used to detect the early and advanced apoptotic cell by flow cytometry, and the expression of apoptosis-associated poteins was investigated by immunohistochemistry or Western-Blot. Results： Stychnine can decrease chondrocyte apoptosis. Meanwhile the early apoptosis ratio of each group was obviously higer than the advanced one and the difference was significant （P（0.05）. The expression of Cyto- chrome C and Smac in cytoplasm of normal group is weak. After SNAP induced chondrocyte apoptosis, the expression of those was obviously higher, and activity decreased as stychnine increasing. Conclusions： Stychnine can decrease chondrocyte apoptosis of SD rat induced by SNAP. The protection function for articular cartilage is likely realized by the mitochondrial pathway.

关 键 词：骨关节炎 软骨细胞 凋亡 线粒体 

分 类 号：R966[医药卫生—药理学] R684.3[医药卫生—药学]